Name: ecm protein nanofibers and nanostructures engineered using surface-initiated assembly
Uploaded: 2014-04-17T16:00:00
Description: Watch this Scientific Journal Video about ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly at JoVE.com

财政支持是来自美国国立卫生研究院主任的新的创新奖（1DP2HL117750）提供给JMS来自美国国立卫生研究院生物力学在再生医学T32训练计划（2T32EB003392），以QJ从多德 - ICES奖学金和AWF。

1，制作母模使用光刻<ol><li>细胞外基质蛋白纳米纤维，nanofabrics和纳米结构是捏造的，首先设计使用计算机辅助设计（CAD）软件。然后这个CAD文件被传送到一个​​光掩模。光罩的类型将取决于功能的决议;用透明度为基础的光罩足够的特征尺寸降低到〜10微米。较小的特点&lt;10微米，需要在玻璃光掩膜铬。所有这里介绍的纳米纤维和纳米结构是使用一个透明的基于光掩模制​​造的，因而是纳?...

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这里的新航法提出模仿细胞介导的​​基质组装，使之ECM蛋白纳米纤维和纳米结构工程与可调大小，组织和组成。虽然不完全相同的细胞产生的细胞外基质，新航造成细胞外基质蛋白纳米纤维20的经历可逆的折叠/内机械应变21展开，并能结合细胞20组成。这提供了构建的ECM蛋白的材料，概括ECM的体内发现了许多性能独特的能力。例如，细胞外基质的纳米纤维可以制造...

细胞外基质（ECM）的组织是由参与多种细胞过程，包括黏附，增殖，分化和凋亡1-3的物理和化学调控的多功能蛋白质。 ECM的合成，组装，并通过细胞组织组成的蛋白质纤维具有独特的成分，纤维尺寸，几何形状和相互关联的架构，随组织类型和发展阶段。最近的工作已经证明，细胞外基质可以提供指导性线索来指导细胞形成工程化组织4，这表明扼要的ECM中的组成和结构方面可能使仿生材料的开发用于组织工程和生物技术的应用。 许多制造方法已开发工程师的聚合物支架，可在组织中模仿的ECM的各方面。例如，静电纺丝和相位组合模式振动性也都表现出以形成纤维的多孔基质具有直径范围从几十微米下降到几十纳米5-7的能力。这两种技术也已经表明，纳米纤维的高度多孔的基质可支持细胞粘附和浸润到支架8。然而，这些方法都在能够建立可能的纤维的几何形状，取向和三维结构的限制。静电通常会产生与支架或者进行定向或随机高度一致的纤维，而相分离产生的支架与随机取向的纤维。也有对这些材料的限制，研究人员通常采用的合成聚合物，如聚（ε-己内酯）8和聚（乳酸-共-乙醇酸）9，即其后涂ECM蛋白，以促进细胞粘附。天然生物聚合物也被使用，包括I型胶原10，明胶11，纤维蛋白原12，脱乙酰壳多糖13，和丝14，但是表示在天然组织中发现的蛋白质的仅一小部分。大多数组织中含有ECM蛋白质和多糖，包括纤连蛋白（FN），层粘连蛋白（LN），IV型胶原和透明质酸，很难或不可能制造用现有的方法的纳米纤维的较大的环境。 为了应对这一挑战，我们一直致力于我们对模仿的方式细胞合成，组装和组织细胞外基质蛋白纤维在其周围的研究工作。而具体原纤维形成过程而变化为不同的细胞外基质蛋白，通常在ECM蛋白质分子的构象变化是由酶或受体介导的相互作用，它公开晦涩自组装站点触发。这里我们使用FN作为一个模型系统，以更好地了解纤维形成过程。简言之，FN同源二聚体结合到通过RGD氨基酸序列在第10的II型整合素受体在细胞表面我重复单元。一旦绑定，则通过整合肌动球蛋白收缩移开，然后展开FN二聚体，露出神秘的自组装基地。这些FN-FN的结合位点的曝光使FN二聚体装配成不溶性纤维权利于细胞表面15。在无细胞系统中的工作已经证明，隐蔽FN-FN的结合位点可以通过使用变性剂16或表面张力在空气-液体-固体界面17-19展开的被揭示。然而，通过这些技术产生的FN纤维被限制在特定的纤维的尺寸和几何形状，并且通常结合到一个表面。 在这里，我们描述了一种方法称为表面引发组装体（SIA）20是利用蛋白质表面的相互作用来创建独立的不溶性纳米纤维，nanofabrics（二维表）和一个或多个ECM蛋白组成（ 图1其它纳米结构克服了这些限制）。在这个Process，ECM蛋白质从溶液中一个紧凑的，球形构象吸附和部分变性（展开）上的图案化，疏水性聚二甲基硅氧烷（PDMS）的邮票。的ECM蛋白，然后在这种状态下转移到热响应聚（N-异丙基丙烯酰胺）（PIPAAm）表面通过微接触印刷22。当用40℃的水的水合的PIPAAm仍是固体，但是当冷却到32℃下其穿过其中​​它变成亲水性的低临界溶解温度（LCST），溶胀用水，然后溶解，释放出组装纳米结构的ECM销的表面上。新航方法提供了具有纳米级精度的尺寸控制。通过控制的关键参数，如组合物中，纤维的几何形状，和架构，它可以概括ECM的体内发现了许多属性和开发先进的支架用于组织工程和生物技术的应用。

<table border="0" cellpadding="0" cellspacing="0" width="651">
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Poly(N-isopropylacrylamide) / PIPAAm</td>
			<td style="width:145px;">Polysciences</td>
			<td style="width:123px;">21458-10</td>
			<td style="width:167px;">40,000 Mw</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Sylgard 184 Silicone kit (PDMS)</td>
			<td style="width:145px;">Dow Corning</td>
			<td style="width:123px;"></td>
			<td style="width:167px;">Mix 10 parts base with 1 part curing agent.&#160;</td>
		</tr>
		<tr height="21">
			<td height="42" style="height:42px;width:216px;">Butanol</td>
			<td style="width:145px;"></td>
			<td style="width:123px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Fibronectin</td>
			<td style="width:145px;">BD biosciences</td>
			<td style="width:123px;">354008</td>
			<td style="width:167px;">Human, 1mg</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Laminin</td>
			<td style="width:145px;">BD biosciences</td>
			<td style="width:123px;">354239</td>
			<td style="width:167px;">Ultrapure, mouse, 1mg</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Negative Photoresist</td>
			<td style="width:145px;">Microchem</td>
			<td style="width:123px;">SU8-2015</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">SU8 Developer</td>
			<td style="width:145px;">Microchem</td>
			<td style="width:123px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="55">
			<td height="55" style="height:55px;width:216px;">Sonicator Branson M3510</td>
			<td style="width:145px;">Branson Ultrasonic Corporation</td>
			<td style="width:123px;">CPN-952-318</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="51">
			<td height="51" style="height:51px;width:216px;">Thinky ARE-250 Mixer</td>
			<td style="width:145px;">Thinky Corporation</td>
			<td style="width:123px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Spincoater</td>
			<td style="width:145px;">Specialty Coating Systems</td>
			<td style="width:123px;">G3P-8</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Glass cover 25mm diameter, No 1.5</td>
			<td style="width:145px;">Fisher Scientific</td>
			<td style="width:123px;">12-545-86</td>
			<td style="width:167px;"></td>
		</tr>
	</tbody>
</table>

ecm protein nanofibers and nanostructures engineered using surface-initiated assembly

细胞外基质（ECM）在组织中被合成，并通过细胞组装以形成一个三维纤维状蛋白质与网络紧密调节纤维的直径，组成和组织。除了 ​​提供结构支撑，ECM的物理和化学性质在多种细胞过程，包括粘附，分化和凋亡中发挥重要作用， 在生物体内时，ECM是由蛋白质中暴露隐秘自组装（原纤维）位点组装。这个过程变化为不同的蛋白质，但纤连蛋白（FN）的原纤维是非常明确的，并作为细胞介导的​​ECM组件的模型系统。具体地，将细胞用对细胞膜整合素受体结合FN二聚体和肌动球蛋白生成的收缩力，以展开并暴露结合位点组装成不溶性纤维。该受体介导的过程使细胞从细胞聚集和组织的ECM来组织SCALES。在这里，我们提出了一个方法称为表面引发组装体（SIA），采用蛋白质表面的相互作用展开ECM蛋白和它们组装成不溶性的纤维，概括细胞介导的​​基质组装。首先，ECM蛋白吸附到疏水性聚二甲基硅氧烷（PDMS）表面，他们部分变性（折叠）和揭露隐秘结合结构域。未折叠的蛋白质，然后转移中良好定义的微和纳米图案通过微接触印刷到热响应聚（N-异丙基丙烯酰胺）（PIPAAm）表面。该PIPAAm的热引发溶解导致最终组装和不溶性的ECM蛋白的纳米纤维和纳米结构具有良好定义的几何形状的释放。复杂的架构是可以通过在用于微接触印刷的PDMS印章定义的工程模式。除了FN，新航过程中可以与层粘连蛋白，纤维蛋白原被使用和胶原蛋白I型和IV创建多组分的ECM nanostrucTures的。因此，SIA，可用于工程ECM蛋白质基材料，具有精确控制的蛋白质组合物，纤维的几何形状和脚手架架构中为了概括ECM的结构和组合物在体内 。

新航能工程的ECM蛋白纳米纤维与精确控制光纤的尺寸。为了证明这一点，FN纳米纤维为50×20微米平面尺寸的阵列，图案到PIPAAm涂盖玻片（ 图2A）。获释后，纤维收缩，因为他们是在一个固有的预应力对PIPAAm面（ 图2B）图案时。新生力量纳米纤维的分析表明它们是单分散的预发行以50.19的平均长度±0.49微米的19.98±0.17微米，宽度（ 图2C）。尽管收缩明显，FN纤维?...

Watch this Scientific Journal Video about ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly at JoVE.com

ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly

A到从单个或多个细胞外基质蛋白得到纳米纤维和纳米结构复杂的方法进行描述。此方法使用蛋白质表面的相互作用与可调的组成和架构，在各种组织工程和生物技术应用创造自由站立蛋白质为基础的材料。

细胞外基质蛋白纳米纤维和纳米结构工程采用表面引发大会

The extracellular matrix (ECM) in tissues is synthesized and assembled by cells to form a 3D fibrillar, protein network with tightly regulated fiber ...

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Visualization of Recombinant DNA and Protein Complexes Using Atomic Force Microscopy

重组DNA和蛋白质复合物，用原子力显微镜的可视化

Atomic force microscopy (AFM) allows for the visualizing of individual proteins, DNA molecules, protein-protein complexes, and DNA-protein complexes. On ...

科研

生物工程

Utilization of Plasmonic and Photonic Crystal Nanostructures for Enhanced Micro- and Nanoparticle Manipulation

为增强微型和纳米操纵利用电浆和光子晶体纳米结构

A method to manipulate the position and orientation of submicron particles nondestructively would be an incredibly useful tool for basic biological ...

Synthesis, Assembly, and Characterization of Monolayer Protected Gold Nanoparticle Films for Protein Monolayer Electrochemistry

单层的合成，装配和表征蛋白质单分子膜的电化学保护的金纳米粒子薄膜

Colloidal gold nanoparticles protected with alkanethiolate ligands called monolayer protected gold clusters (MPCs) are synthesized and subsequently ...

Engineered Vascularized Muscle Flap

工程化血管肌瓣

One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen ...

Micropatterning and Assembly of 3D Microvessels

缩微和3D微血管大会

In vitro platforms to study endothelial cells and vascular biology are largely limited to 2D endothelial cell culture, flow chambers with polymer or glass ...

Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method

工程血管移植缩放使用3D打印指南和环堆叠方法

Coronary artery disease remains a leading cause of death, affecting millions of Americans. With the lack of autologous vascular grafts available, ...

Fabrication of Custom Agarose Wells for Cell Seeding and Tissue Ring Self-assembly Using 3D-Printed Molds

用 3 d-印刷模具制作细胞播种和组织环自组装用的自定义琼脂糖井

Engineered tissues are being used clinically for tissue repair and replacement, and are being developed as tools for drug screening and human disease ...

Nondestructive Monitoring of Degradable Scaffold-Based Tissue-Engineered Blood Vessel Development Using Optical Coherence Tomography

基于光学相干断层扫描的可降解支架组织工程血管发育的无损监测

Engineered vascular grafts with structural and mechanical properties similar to natural blood vessels are expected to meet the growing demand for arterial ...

Tissue-Engineered Graft for Circumferential Esophageal Reconstruction in Rats

用于大鼠圆周食管重建的组织工程移植

The use of biocompatible materials for circumferential esophageal reconstruction is a technically challenging task in rats and requires an optimal implant ...

Self-Assembly of Gamma-Modified Peptide Nucleic Acids into Complex Nanostructures in Organic Solvent Mixtures

将伽马改性肽核酸自组装成有机溶剂混合物中复杂的纳米结构

Current strategies in DNA and RNA nanotechnology enable the self-assembly of a variety of nucleic acid nanostructures in aqueous or substantially hydrated ...